Large-Scale Identification of Protein Crotonylation Reveals Its Role in Multiple Cellular Functions

• Published in: Journal of proteome research Vol. 16; no. 4; pp. 1743 - 1752
• Main Authors: Wei, Wei, Mao, Anqi, Tang, Bin, Zeng, Qiufang, Gao, Shennan, Liu, Xiaoguang, Lu, Lu, Li, Wenpeng, Du, James X, Li, Jiwen, Wong, Jiemin, Liao, Lujian
• Format: Journal Article
• Language: English
• Published: United States American Chemical Society 07.04.2017
• Subjects: Acyl Coenzyme A - metabolism; Cell Cycle Proteins - isolation & purification; Cell Cycle Proteins - metabolism; DNA Replication - genetics; HeLa Cells; Histones - metabolism; Humans; Lysine - metabolism; Mass Spectrometry - methods; Promoter Regions, Genetic; Protein Processing, Post-Translational - genetics; Proteomics; mass spectrometry; DNA replication; lysine crotonylation; proteomics; cell cycle
• ISSN: 1535-3893; 1535-3907
• DOI: 10.1021/acs.jproteome.7b00012

Lysine crotonylation on histones is a recently identified post-translational modification that has been demonstrated to associate with active promoters and to directly stimulate transcription. Given that crotonyl-CoA is essential for the acyl transfer reaction and it is a metabolic intermediate widely localized within the cell, we postulate that lysine crotonylation on nonhistone proteins could also widely exist. Using specific antibody enrichment followed by high-resolution mass spectrometry analysis, we identified hundreds of crotonylated proteins and lysine residues. Bioinformatics analysis reveals that crotonylated proteins are particularly enriched for nuclear proteins involved in RNA processing, nucleic acid metabolism, chromosome organization, and gene expression. Furthermore, we demonstrate that crotonylation regulates HDAC1 activity, expels HP1α from heterochromatin, and inhibits cell cycle progression through S-phase. Our data thus indicate that lysine crotonylation could occur in a large number of proteins and could have important regulatory roles in multiple nuclei-related cellular processes.